A sensitive and specific enzyme immunoassay for the detection of methyl ether derivatives of cyclomaltoheptaose

Abstract

We have raised antibodies against two methylated derivatives of β-CD, heptakis(2,6-di- O-methyl)cyclomaltoheptaose (Dimeb) and heptakis(2,3,6-tri- O-methyl)cyclomaltoheptaose (Trimeb). These antibodies were used to develop two specific and sensitive enzyme immunoassays, presenting a detection limit close to 500 and 30 pg/mL for Trimeb and Dimeb, respectively. Cross reactivities of different linear and cyclic maltooligosaccharides were investigated, demonstrating a high specificity against the structural features of the secondary hydroxyls rim. Several commercial Dimeb samples, containing different mixtures of partially methylated β-cyclodextrin derivatives including RAMEB, which contains only a few amount of pure Dimeb, could be easily evaluated by the Dimeb immunoassay. Both of these assays have been shown to allow accurate measurement in plasma and urine, thus appearing as useful tools for further applications in biological material.